Table S1. Strains and plasmids used in this study

Strain or plasmid / Relevant genotype or phenotype / Source or reference
Strain
E. coli
TOP10 / F-mcrA, D(mrr-hsdRMS-mcrBC), f80lacZDM15, DlacX74, recA1, deoR, araD139, D(ara-leu)7679, galU, galK, rpsL (StrR), endA1, nupG / Invitrogen
S17-1λpir / recA, thi, pro, hsdR-M+, SmR, <RP4:2-Tc:Mu:Ku:Tn7>TpR / [73]
DH5αF¢IQ / F-φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk-, mk+) phoA supE44 λ- thi-1 gyrA96 relA1/F´ proAB+ lacIqZΔM15 zzf::Tn5 [KmR]. / Invitrogen
KDZif1ΔZ / B2H reporter strain, KmR, CmlR / [74]
F. tularensis
LVS / Live vaccine strain / USAMRIID1
DiglA / LVS, iglA in-frame deletion of codons 4-174 / [14]
DiglB / LVS, iglB in-frame deletion of codons 54-346 / [14]
DiglC / LVS, iglC in-frame deletion of codons 28-205 / [75]
DiglD / LVS, iglD in-frame deletion mutant / [68]
DdotU / LVS, dotU in-frame deletion of codons 4-203 / This study
DvgrG / LVS, vgrG in-frame deletion of codons 4-162 / This study
DvgrG/VgrGcis / DvgrG, in cis complementation of one vgrG deletion copy / This study
S. cerevisiae
AH109 / MATα, trp1-901, leu2-3, 112, ura3-52, his3-200, gal4D, gal80D, LYS2::GAL1UAS-GAL1TATA-HIS3, GAL2UASGAL2TATA-ADE2, URA3::MEL1UAS-MEL1TATA-lacZ, MEL1 / Clontech Laboratories
Plasmid
pCR®4-TOPO® / TA cloning vector, KmR, CbR / Invitrogen
pDM4 / Suicide plasmid carrying sacBR, CmR / [76]
pMOL72 / pDM4 carrying a XhoI/SacI PCR fragment of DvgrG4-162 with flanking regions, CmR / This study
pJEB752 / pDM4 carrying a XhoI/SacI PCR fragment of DdotU4-203 with flanking regions, CmR / This study
pJEB926 / pDM4 carrying a XhoI/SacI PCR fragment of vgrG with flanking regions, CmR / This study
pKK289Km / expression plasmid carrying a gfp gene under the control of the LVS groE promoter, KmR / [68]
pMOL128 / pKK289Km encoding DotU, KmR / This study
pMOL71 / pKK289Km encoding VgrG, KmR / This study
pJEB390 / pKK289Km encoding IglA-GSK, KmR / This study
pJEB391 / pKK289Km encoding IglC-GSK, KmR / This study
pMOL52 / pKK289Km derivative used to construct C-terminal fusion proteins to eukaryotic GSK, KmR / [23]
pMOL58 / pMOL52 encoding DotU-GSK, KmR / This study
pJEB914 / pMOL52 encoding DotU DE70-71AA-GSK, KmR / This study
pJEB915 / pMOL52 encoding DotU DE70-71KK-GSK, KmR / This study
pJEB916 / pMOL52 encoding DotU DE70-71SS-GSK, KmR / This study
pJEB917 / pMOL52 encoding DotU G134A-GSK, KmR / This study
pJEB918 / pMOL52 encoding DotU G134S-GSK, KmR / This study
pJEB919 / pMOL52 encoding DotU G134K-GSK, KmR / This study
pJEB920 / pMOL52 encoding DotU G134D-GSK, KmR / This study
pMOL54 / pMOL52 encoding VgrG-GSK, KmR / This study
pJEB702 / pMOL52 encoding IglB-GSK, KmR / This study
pJEB703 / pMOL52 encoding IglD-GSK, KmR / This study
pACTR-AP-Zif / B2H vector, directs the synthesis of a Zif268-DNA binding domain fusion protein, TetR / [74]
pACTR-MglA-Zif / pACTR-AP-Zif encoding MglA, TetR / [61]
pJEB871 / pACTR-AP-Zif encoding DotU, TetR / This study
pJEB873 / pACTR-AP-Zif encoding VgrG, TetR / This study
pJEB876 / pACTR-AP-Zif encoding IcmF, TetR / This study
pMOL135 / pACTR-AP-Zif encoding IglA, TetR / This study
pMOL139 / pACTR-AP-Zif encoding IglB, TetR / This study
pLM5 / pACTR-AP-Zif encoding IglC, TetR / This study
pBRGPω / B2H vector, directs the synthesis of a Gal11P-ω fusion protein, CbR / [74]
pBRSspA-ω / pBRGPω encoding SspA, CbR / [61]
pJEB872 / pBRGPω encoding DotU, CbR / This study
pJEB874 / pBRGPω encoding VgrG, CbR / This study
pJEB877 / pBRGPω encoding IcmF, CbR / This study
pMOL133 / pBRGPω encoding IglA, CbR / This study
pMOL134 / pBRGPω encoding IglB, CbR / This study
pLM6 / pBRGPω encoding IglC, CbR / This study
pGADT7 / Y2H vector, LEU2, CbR / Clontech Laboratories
pMOL47 / pGADT7 encoding DotU, LEU2, CbR / This study
pMOL31 / pGADT7 encoding VgrG, LEU2, CbR / This study
pJEB545 / pGADT7 encoding IcmF363-1093, LEU2, CbR / [14]
pJEB890 / pGADT7 encoding IcmF, LEU2, CbR / This study
pJEB393 / pGADT7 encoding IglA, LEU2, CbR / [14]
pJEB395 / pGADT7 encoding IglB, LEU2, CbR / [14]
pJEB397 / pGADT7 encoding IglC, LEU2, CbR / [14]
pGBKT7 / Y2H vector, TRP1, KmR / Clontech Laboratories
pMOL51 / pGBKT7 encoding DotU, TRP1, KmR / This study
pMOL37 / pGBKT7 encoding VgrG, TRP1, KmR / This study
pJEB546 / pGBKT7 encoding IcmF363-1093, TRP1, KmR / [14]
pJEB891 / pGBKT7 encoding IcmF, TRP1, KmR / This study
pJEB392 / pGBKT7 encoding IglA, TRP1, KmR / [14]
pJEB394 / pGBKT7 encoding IglB, TRP1, KmR / [14]
pJEB396 / pGBKT7 encoding IglC, TRP1, KmR / [14]

1 US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD.